技术资料

Classical intracellular delivery methods such as transfection and transduction are inefficient,particularly with confluent cells and organoids,and lack cell type‐specific programmability. We demonstrate that an innovative methodology called calcium shock (CaSh) dramatically improves particle delivery into single cells,colonies,and organoids,and enables programmable delivery (CaSh‐Pro) into specific cell types within heterocellular populations. Calcium shock works by increasing endocytotic uptake while simultaneously disarming cell‐cell junctions. CaSh‐Pro further incorporates specific molecular targeting agents and amphiphilic peptides for preferential editing of different cell types. Calcium shock improves expression of plasmid,ribonucleoprotein,or adeno‐associated viral vectors with minimal toxicity in intact organoids representing diverse lineages. CaSh and CaSh‐Pro provide simple,versatile protocols for genome editing in complex systems,to enable biological discovery and therapeutic development. Classical transfection and transduction are inefficient,particularly with confluent cells and organoids,and lack cell type‐specific programmability. This study presents calcium shock (CaSh),a method that dramatically improves particle delivery into single cells,colonies,and organoids. CaSh is further utilized to enable programmable delivery (CaSh‐Pro) into specific cell types within heterocellular populations. View Publication